Ifnγ pe cy7 xmg1

Manufactured by BioLegend

Sourced in United States

IFNγ-PE/Cy7 (XMG1.2) is a fluorochrome-conjugated antibody that binds to the interferon-gamma (IFNγ) protein. It can be used for the identification and quantification of IFNγ-producing cells in flow cytometry applications.

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2 protocols using ifnγ pe cy7 xmg1

Murine Immune Response Analysis

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Lymph nodes and spleen tissues were collected after mice were euthanatized on day 30. Single cells were obtained by grinding tissues with 70 μm nylon filters in 0.5% BSA-PBS. Anti-mouse antibodies against CD3-Percp/Cy5.5 (145-2C11, BioLegend), CD4-APC (GK1.5, BioLegend), CD4-FITC (GK1.5, BioLegend), CD8-PE (53-6.7, BioLegend), CD11c-FITC (N418, BioLegend), CD11b-Percp/Cy5.5 (M1/70, BioLegend), CD80-PE (16-10A1, eBioscience), CD86-APC (24F, BioLegend), CD69-FITC (H1.2F3, BioLegend), B220-Percp/Cy5.5 (RA3-6B2, BioLegend), CD138-APC (281-2, BioLegend) or I-A/I-E (MHC II)-PE/Cy7 (M5/114.15.2, BioLegend) were used for cell surface antigen staining. After washing with PBS for three times, cells were treated with Foxp3/transcription factor fixation/permeabilization concentrate and diluent (eBioscience, USA) and incubated with anti-mouse antibodies against IFNγ-PE/Cy7 (XMG1.2, BioLegend) or Ki67-FITC (SolA15, BioLegend) for intracellular antigen staining. Besides, S1 protein with His tag (Sino Biological, China) and anti-His tag-PE (BioLegend) were used to label S1 specific B cells. The cells were analyzed by flow cytometry.

Zhang M., Wang L., Liu J, & Pang Y. (2022). Envelope virus-mimetic nanovaccines by hybridizing bioengineered cell membranes with bacterial vesicles. iScience, 25(6), 104490.

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Hybridoma Cell Culture and Antibody Purification

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Hybridoma cell line producing H35 was grown in IMDM culture medium supplemented with 2% of IgG-depleted FCS, 10 mM HEPES, and 50 μM β-mercaptoethanol at 37°C in 5% CO2 atmosphere. The anti-CD8β mAb was purified from cell culture supernatant of the H35 hybridoma on Protein G sepharose column (Amersham). Before immuno-staining, dead cells were excluded with the fixable viability dye eFluor506 (eBioscience). All antibodies used for flow cytometry experiments were diluted in an anti-mouse Fc receptor mAb purified from the supernatant of culture of the 2.4G2 hybridoma. Fluorochrome-conjugated monoclonal antibodies for flow cytometry were purchased from eBioscience: CD3-eFluor450 (eBio500A2), CD11c-PECy7 (N418), CD8α PerCpCy5.5 (53-6.7), CD62L Alexa Fluor700 (MEL-14), CD44 APC (IM7), and IFNγ PECy7 (XMG1.2) or BioLegend: CD19 PE (6D5) and CD4 APCCy7 (RM4-5). Analyses were performed with the FACSLSRII or FACSCanto machines (Becton Dickinson) using the FACSDiva software for the acquisition and the FlowJo software for data analyses.

Duval A., Fuertes Marraco S.A., Schwitter D., Leuenberger L, & Acha-Orbea H. (2014). Large T Antigen-Specific Cytotoxic T Cells Protect Against Dendritic Cell Tumors through Perforin-Mediated Mechanisms Independent of CD4 T Cell Help. Frontiers in Immunology, 5, 338.

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